Split aptamer regulated CRISPR/Cas12a biosensor for 17β-estradiol through a gap-enhanced Raman tags based lateral ﬂow strategy

Qing Li; Xiaobo Li; Pengyou Zhou; Rui Chen; Rui Xiao; Yuanfeng Pang

doi:10.1016/j.bios.2022.114548

Split aptamer regulated CRISPR/Cas12a biosensor for 17β-estradiol through a gap-enhanced Raman tags based lateral ﬂow strategy

Biosens Bioelectron. 2022 Nov 1:215:114548. doi: 10.1016/j.bios.2022.114548. Epub 2022 Jul 12.

Authors

Qing Li¹, Xiaobo Li¹, Pengyou Zhou¹, Rui Chen², Rui Xiao³, Yuanfeng Pang⁴

Affiliations

¹ Capital Medical University, Department of Toxicology, No. 10 Xitoutiao, You An Men, Beijing, 100069, PR China.
² Capital Medical University, Department of Toxicology, No. 10 Xitoutiao, You An Men, Beijing, 100069, PR China. Electronic address: ruichen@ccmu.edu.cn.
³ Beijing Institute of Microbiology and Epidemiology, 27 Taiping Road, 100850, Beijing, PR China. Electronic address: ruixiao203@sina.com.
⁴ Capital Medical University, Department of Toxicology, No. 10 Xitoutiao, You An Men, Beijing, 100069, PR China. Electronic address: yuanfengpang@ccmu.edu.cn.

PMID: 35870335
DOI: 10.1016/j.bios.2022.114548

Abstract

It is significant to exploit the full potential of CRISPR/Cas based biosensor for non-nucleic-acid targets. Here, we developed a split aptamer regulated CRISPR/Cas12a and gap-enhanced Raman tags based lateral ﬂow biosensor for small-molecule target, 17β-estradiol. In this assay, one split aptamer of 17β-estradiol was designed to complement with crRNA of Cas12a so that the trans-cleavage ability of CRISPR/Cas12a can be regulated by the competitive binding of 17β-estradiol and split aptamers. Through integration of the signal amplification ability of CRISPR/Cas12a and the ultra-sensitive gap-enhanced Raman tags based lateral ﬂow assay, a visible-SERS dual mode determination of 17β-estradiol can be established. 17β-estradiol can be visibly recognized as low as 10 pM and accurately quantified with a detection limit of 180 fM by SERS signals, which is at least 10³-fold lower than that of the previous immunoassay lateral ﬂow strategies. Our assay provides a novel perspective to develop split aptamer regulated CRISPR/Cas12a coupling with SERS lateral ﬂow strips for ultrasensitive and easy-to-use non-nucleic-acid targets detection.

Keywords: 17β-estradiol; CRISPR/Cas12a; Gap-enhanced Raman tags; Lateral flow assay; Split aptamers.

MeSH terms

Aptamers, Nucleotide* / chemistry
Biosensing Techniques*
CRISPR-Cas Systems / genetics
Estradiol / chemistry

Substances

Aptamers, Nucleotide
Estradiol